STANDARD OPERATING PROCEDURE

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Ref.. Table of Contents Page. Scope 2 2. Objective 2 3. Definitions 2 4. Responsibility 3 5. Procedure 3 5.. Diagram of Display and Control Panel 3 5.2. Installation 4 5.3. Operation 4 5.4. Calibration 5 5.5. Maintenance 6 5.6. Troubleshooting 7 5.7. Flow charts 8 6. Precautions 3 7. References 3 8. Appendices 3 9. Revision History 4 Page of 4

Ref... Scope This Standard Operating Procedure (SOP) applies to the staff and students making use of the Hanna Portable ph Meter HI 9025 in the laboratories of the Pharmacy Department, University of Malta. 2. Objective To describe the procedure for the installation, operation, calibration, maintenance and troubleshooting of the Hanna Portable ph Meter HI 9025. 3. Definitions 3.. o C and o C Keys (8): To manually set the temperature or the ph buffer value. 3.2. Automatic Temperature Compensation (ATC): A feature in which the temperature probe is able to detect the temperature of the solution in which it is immersed in. 3.3. Bayonet Neill-Concelman (BNC) Electrode connector (): A common type of cable connector used to connect the ph electrode to the meter. 3.4. CAL Key (6): To enter or exit calibration mode. 3.5. CFM Key (7): To confirm calibration. 3.6. Display (3): Digitally displays the ph, temperature and mv results. 3.7. MEM Key (9): To store a ph value in memory. 3.8. MR Key (0): To recall the stored value from memory. 3.9. Manual Temperature Compensation: A feature in which the temperature of the sample solution is manually inputted when this is already known. 3.0. On/Off Key (5): To turn the meter on or off. Page 2 of 4

Ref.. 3.. Primary Display (): The upper part of the display that shows the resultant ph, temperature and mv value readings. 3.2. RANGE Key (4): To select between ph and mv results. 3.3. Secondary Display (2): The lower part of the display that shows a number of different commands to be taken. 3.4. Temperature Probe Socket (2): The socket at which the temperature probe is connected to the meter. 4. Responsibility 4.. The members of the Department of Pharmacy (staff and students) are responsible for following this SOP. 4.2. The designated Laboratory Officer or Laboratory Assistant is responsible for ensuring that this SOP is followed. 5. Procedure 5.. Diagram of Display and Control Panel 2 Page 3 of 4

Ref.. 5.2. Installation 5.2.. Install batteries in the back compartment of the meter. 5.2.2. Connect the ph electrode and the temperature probe to the BNC and temperature sockets respectively. 5.2.3. Switch the meter on by pressing and holding the On/Off Key for a fraction of a second. 5.3. Operation 5.3.. Measuring ph 5.3... Ensure that before use, the meter is being calibrated frequently to allow more accurate measurements to be taken. 5.3..2. Remove the electrode protective cap. 5.3..3. Submerge the ph electrode into the sample to be tested. 5.3..4. Turn the meter on and if necessary press the RANGE Key until the display changes to the ph mode. 5.3..5. Allow the ph electrode to adjust and stabilise in the sample. 5.3..6. Read off the displayed stable reading. 5.3.2. Automatic and Manual Temperature Compensation 5.3.2.. Submerge the temperature probe into the sample as close to the ph electrode as possible and wait for a couple of minutes to allow it to stabilise if automatic temperature compensation is desired. 5.3.2.2. Disconnect the temperature probe from the meter if manual temperature compensation is desired whenever the temperature of the sample to be tested is already known. Adjust the displayed temperature by using the Up and Down Keys respectively. 5.3.3. Measuring Temperature 5.3.3.. Turn the meter on and press the RANGE Key until the display changes to temperature mode. 5.3.3.2. Dip the temperature probe into the sample. Page 4 of 4

Ref.. 5.4. Calibration 5.3.3.3. Allow a couple of minutes for the reading to stabilise. 5.3.3.4. Read off the displayed stable reading. 5.4.. Pour small quantities of ph 7.0 and ph 4.0 or ph 0.0 buffer solutions into two separate clean beakers and use another two separate beakers for rinsing. 5.4.2. Switch on meter and ensure that it is in ph mode. 5.4.3. Press the CAL Key. 5.4.4. Ensure that the secondary display is showing the desired buffer solution and if not, press the Up and Down keys until the desired ph of the buffer solution to be used is displayed. 5.4.5. Remove protective cap and rinse the electrode with the ph 7.0 buffer calibration solution. 5.4.6. Immerse the ph electrode into the ph 7.0 buffer solution and stir gently. 5.4.7. Ensure that the electrode is kept submerged approximately 4cm into the solution. 5.4.8. Ensure that the temperature probe is located as close to the ph electrode as possible. 5.4.9. Wait until the display has [READY] and [CON] blinking to ensure that reading is stable. 5.4.0. Press the CFM Key to confirm the calibration. 5.4.. If [WRONG] is seen blinking, ensure that the buffer solution used corresponds with the buffer solution chosen in the meter. 5.4.2. Wait until the ph of the buffer is displayed on the primary display and the ph of another buffer solution is being displayed on the secondary display. 5.4.3. Press the Up and Down keys to select the desired ph buffer solution (ph 4.0 or ph 0.0) for the next calibration. 5.4.4. Rinse the ph electrode with distilled water and then with the second buffer solution. 5.4.5. Immerse the ph electrode into the second buffer solution and stir gently. 5.4.6. Wait until the display has [READY] and [CON] blinking to ensure that reading is stable. 5.4.7. Press the CFM Key to confirm the calibration. 5.4.8. If [WRONG] is seen blinking, ensure that the buffer solution used corresponds with the buffer solution chosen in the meter. Page 5 of 4

Ref.. 5.4.9. Wait until meter returns to the operating mode to ensure that calibration process was successful. 5.5. Maintenance 5.5.. Inspect the cables of the electrodes for any signs of broken insulation. 5.5.2. Inspect the ph electrode for any cracks in its stem or bulb. 5.5.3. Rinse off any salt desposits with water. 5.5.4. Soak the ph electrode in Hanna HI 706 or HI 806 General Cleaning Solution for approximately ½ hour to perform general cleaning. 5.5.5. Soak the ph electrode in Hanna HI 7073 or HI 8073 Protein Cleaning Solution for 5 minutes if protein deposits are present on the membrane and/or junction of the electrode. 5.5.6. Soak the ph electrode in Hanna HI 7074 or HI 8074 Inorganic Cleaning Solution for 5 minutes if inorganic deposits are present on the membrane and/or junction of the electrode. 5.5.7. Rinse the ph electrode with Hanna HI 7077 or HI 8077 Oil and Fat Cleaning Solution if oil films or deposits are present on the membrane and/or junction of the electrode. 5.5.8. Rinse the ph electrode thoroughly with distilled water after performing any of these cleaning procedures. 5.5.9. Refill the reference chamber with fresh electrolyte and soak it in HI 70300 or HI 80300 Storage Solution for at least hour before taking any measurements. 5.5.0. If [LOW BAT] is displayed on the lower left hand corner of the secondary display, unscrew the back panel of the meter, remove all 4 batteries and replace with new.5v ones ensuring that their polarity is correct. Page 6 of 4

Ref.. 5.6. Trouble Shooting Fault Possible Cause/s Solution Electrode not working or reference junction clogged Meter working slowly or giving faulty readings Meter dose not accept 2 nd buffer solution for calibration Reading is drifting Display shows [E] or no ph value ph electrode is out of order Leave electrode in storage solution after cleaning junction. If problem persists, replace electrode Follow normal cleaning procedure. If problem persists, replace electrode ph electrode is out of Replace ph electrode order ph scale is out of range - Recalibrate - Ensure ph sample is in the 0 to 4 range - Check electrolyte level and the general state of the ph electrode Display shows [E2] or no ph and temperature values Display shows [E3] or no mv value Display shows [E4] or [WRONG BUF ] Meter is not working with temperature probe Meter fails to calibrate or gives faulty readings Display is acting erratically Temperature scale is out of range mv scale is out of range - Erroneous buffer solution used for offset calibration - Out of order electrode Temperature probe is out of order ph electrode is out of order Microprocessor electronically distrubed - Ensure temperature probe is plugged in - Ensure temperature is in the 0 to 00 o C range Appropriate electrode is not connected - Ensure buffer setting in meter is correct and the solution used is fresh - Replace the ph electrode Replace temperature probe Replace ph electrode Remove one of the batteries for minute to reset microprocessor Page 7 of 4

Ref.. 5.7. Flow Charts 5.7.. Installation Start Install batteries in back compartment of meter Connect ph electrode and T probe to BNC and T sockets respectively Switch meter on by pressing On/Off key for a fraction of a second End Page 8 of 4

Ref.. 5.7.2. Operation Start Measure ph Measure T Calibration carried out frequently Remove electrode protective cap Perform calibration (see section 5.7.3) Turn meter on Display in T mode Press RANGE Key until display changes to T mode Dip T probe into sample Submerge ph electrode into sample to be tested Turn meter on Wait for a couple of minutes to allow it to stabilise Display in ph mode Press RANGE Key until display changes to ph mode Read off displayed stable T reading End Allow ph electrode to adjust and stabilise in sample Use ATC feature MTC Attach T probe to T socket Change displayed T with T of sample by using the Up and Down Keys Submerge T probe into sample as close to ph electrode as possible Wait for a couple of minutes to allow it to stabilise Read off displayed stable ph reading Page 9 of 4

Ref.. 5.7.3. Calibration Start Pour small quantities of ph 7.0 and ph 4.0 or ph 0.0 buffer solutions into 2 separate clean beakers Use 2 other beakers for rinsing Switch on meter Meter in ph mode Press RANGE Key until display changes to ph mode Press CAL Key 2 o display showing desired buffer solution Press Up and Down keys until desired buffer solution is displayed 2 Remove protective cap Rinse electrode with some of the ph 7.0 buffer solution Immerse electrode into ph 7.0 buffer solution and stir gently Electrode submerged 4cm into solution Submerge accordingly Place the T probe as close to the ph electrode as possible Page 0 of 4

Ref.. [WRONG] seen blinking Wait until [READY] and [CON] are blinking on display Press CFM Key to confirm calibration Buffer solution used corresponds with buffer solution in meter Arrange accordingly Wait until ph of buffer is displayed on o display Use a fresh supply of buffer solution 2 Desired ph of next buffer solution displayed on 2 o display Press Up and Down keys until desired buffer solution is displayed 2 3 Rinse ph electrode with distilled water and then rinse with second buffer solution Immerse ph electrode into this second buffer solution and stir gently [WRONG] seen blinking Wait until [READY] and [CON] are blinking on display Press CFM Key to confirm calibration Arrange accordingly Buffer solution used corresponds with buffer solution in meter Wait until programme returns to operating mode to ensure calibration was successful 3 End Use a fresh supply of buffer solution 3 Page of 4

Ref.. 5.7.4. Maintenance Start Inspect cables of electrodes for any signs of broken insulation Inspect ph electrode for any cracks in stem or bulb Rinse off any salt deposits with water General cleaning Protein deposits present Oil films / deposits present Soak ph electrode in Hanna HI 706 or HI 806 General Cleaning Solution for ½ hour Soak ph electrode in Hanna HI 7073 or HI 8073 Protein Cleaning Solution for 5 minutes Soak ph electrode in Hanna HI 7074 or HI 8074 Inorganic Cleaning Solution for 5 minutes Inorganic deposits present 2 Soak ph electrode in Hanna HI 7074 or HI 8074 Inorganic Cleaning Solution for 5 minutes Rinse ph electrode thoroughly with distilled water Refill reference chamber with fresh electrolyte 2 Soak in HI 70300 or HI 80300 Storage Solution for at least hr before use [LOW BAT] displayed Unscrew back panel of meter, remove all 4 batteries and replace with new.5v ones ensuring polarity is correct Page 2 of 4 End

Ref.. 6. Precautions 6.. Always keep the ph electrode wet and rinse it thoroughly with the sample to be measured before use. 6.2. Keep in mind that the ph is directly affected by temperature. Thus in order to measure the ph accurately, the temperature must be taken into consideration. 6.3. If the sample temperature is quite different from the temperature at which the ph electrode is kept, allow a few minutes for thermal equilibrium to take place. 6.4. When performing a two-point calibration, use ph 7.0 and ph 4.0 when measuring acidic samples and ph 7.0 and ph 0.0 for alkaline measurements. This provides more accurate readings to be taken. 6.5. Do not be alarmed when salt deposits are present on the electrode since these normally form and can easily be rinsed off with water. 6.6. If any air bubbles form inside the glass bulb, shake it down firmly to remove them. 6.7. Always store the electrode in its appropriate storage solution and never store it in distilled or deionised water. 6.8. Perform a calibration: 7. References 6.8.. When ph electrode or temperature probe is replaced 6.8.2. At least once a month 6.8.3. After testing aggressive chemicals 6.8.4. When batteries have been replaced 6.8.5. When greater accuracy is required Hanna Instruments. Instruction Manual HI 9023C HI 9023 CN HI 9024C HI 9025C HI 90 HI 920 Portable Waterproof ph Meters. Bedfordshire: Hanna Instruments; 2000. 8. Appendices N/A Page 3 of 4

Ref.. 9. Revision History Version Number Amendments/ Reasons for change 0 Initial Release 02 Inclusion of subheading titles of Procedure section in Table of Contents Page 4 of 4